These days I’ve been detecting clearer bands in the centre of the membrane while the bands on the sides are ok... Even with the actins or GAPDH..!! Now I can't rely on my results... Does anybody know why is this happening to me?

I had this problem for a while, and it was because my sandwiches were packed too tightly and were just "squeezed" better on the ends resulting in a more efficient transfer at the edges. Did you recently switch filter papers or sponges? That's when we caught it.

Are you using a stir bar in your transfer? That can be another source of what you're seeing - too much turbulence in the center. I no longer use the stir bar and have no problems with overheating.

Best of luck.

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